| 重组毕赤酵母表达瑞替普酶(Reteplase)过程中的降解控制 |
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| 引用本文： | 邹敏,方曙光,黄立,庄英萍,储炬,张嗣良,严成钊.重组毕赤酵母表达瑞替普酶(Reteplase)过程中的降解控制[J].工业微生物,2006,36(2):7-11. |
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| 作者姓名： | 邹敏 方曙光 黄立 庄英萍 储炬 张嗣良 严成钊 |
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| 作者单位： | 1. 华东理工大学生物反应器工程国家重点实验室,上海200237
2. 上海市工业微生物研究所,上海,200233 |
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| 基金项目： | 国家高技术研究发展计划“863计划”(No.2002AA217021,2002AA2Z3451) |
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| 摘 要： | 在利用重组毕赤酵母表达瑞替普酶(Reteplase)时,遇到了较为严重的蛋白降解。为了抑制Reteplase的降解,分别研究了pH和氨基酸、蛋白胨等添加物对降解的影响。结果表明,摇瓶培养时用氨水调节pH,并将pH恒定在6.5能有效地抑制降解,这一结果在5L罐上也得到了验证,表达量提高到487mg/L。添加0.1%的Leu或Ala和将蛋白胨用量增加至6%时,Reteplase的降解也能得到有效的控制。
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| 关 键 词： | pH 氨基酸 蛋白胨 毕赤酵母 rPA |
Control of reteplase degradation during expression process by Pichia pastoris |
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| ZOU Min,FANG Shu-guang,HUANG Li,ZHUANG Ying-ping,CHU Ju,ZHANG Si-liang,YAN Cheng-zhao.Control of reteplase degradation during expression process by Pichia pastoris[J].Industrial Microbiology,2006,36(2):7-11. |
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| Authors: | ZOU Min FANG Shu-guang HUANG Li ZHUANG Ying-ping CHU Ju ZHANG Si-liang YAN Cheng-zhao |
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| Institution: | 1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237 ; 2. Shanghai Institute of Industrial Microbiology, Shanghai 200233 |
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| Abstract: | Severe degradation of reteplase expressed by Pichia pastoris was observed.Effects of pH and addition of amino acids and polypeptone were investigated in order to minimize the degradation of target protein.In shake flasks,it was suggested that rPA degradation could be alleviated effectively at pH 6.5 when ammonia was used to adjust the pH.When the results were applied to 5L fermentor,the concentration of rPA was increased to 487mg/L.At the same time,the addition of(0.1%) Leu or Ala and the increasement of polypeptone content to 6% in the medium could also reduce rPA degradation to some extent. |
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| Keywords: | pH amino acid polypeptone Pichia pastoris rPA |
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